Repetitive lagging strand DNA synthesis by the bacteriophage T4 replisome.
Our studies on the T4 replisome build on the seminal work from the Alberts laboratory. They discovered essentially all the proteins that constitute the T4 replisome, isolated them, and measured their enzymatic activities. Ultimately, in brilliant experiments they reconstituted in vitro a functioning replisome and in the absence of structural information created a mosaic as to how such a machine might be assembled. Their consideration of the problem of continuous leading strand synthesis opposing discontinuous lagging strand synthesis led to their imaginative proposal of the trombone model, an illustration that graces all textbooks of biochemistry. Our subsequent work deepens their findings through experiments that focus on defining the kinetics, structural elements, and protein-protein contacts essential for replisome assembly and function. In this highlight we address when Okazaki primer synthesis is initiated and how the primer is captured by a recycling lagging strand polymerase--problems that the Alberts laboratory likewise found mysterious and significant for all replisomes.
Spiering
Michelle M
MM
Department of Chemistry, The Pennsylvania State University, University Park, PA 16802, USA.
Nelson
Scott W
SW
Benkovic
Stephen J
SJ
D017122:Bacteriophage T4 / Q000201:enzymology / Q000235:genetics* / Q000378:metabolism; D004247:DNA / Q000096:biosynthesis* / Q000737:chemistry / Q000378:metabolism; D004261:DNA Replication; D004259:DNA-Directed DNA Polymerase / Q000737:chemistry / Q000235:genetics / Q000378:metabolism*; D008954:Models, Biological; D009097:Multienzyme Complexes / Q000737:chemistry / Q000378:metabolism*; D018741:Replication Origin; D013698:Templates, Genetic; D000099272:Replisomes
D004247:DNA; D004259:DNA-Directed DNA Polymerase; D009097:Multienzyme Complexes; D000099272:Replisomes; C022262:Okazaki fragments